Changes in Taste Neurons Support the Emergence of an Adaptive

Behavior in Cockroaches
Ayako Wada-Katsumata et al.
Science 340, 972 (2013);
DOI: 10.1126/science.1234854

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REPORTS

Changes in Taste Neurons Support

the Emergence of an Adaptive
Behavior in Cockroaches
Ayako Wada-Katsumata, Jules Silverman, Coby Schal*
In response to the anthropogenic assault of toxic baits, populations of the German cockroach have rapidly
evolved an adaptive behavioral aversion to glucose (a phagostimulant component of baits). We hypothesized
that changes in the peripheral gustatory system are responsible for glucose aversion. In both wild-type and
glucose-averse (GA) cockroaches, D-fructose and D-glucose stimulated sugargustatory receptor neurons
(GRNs), whereas the deterrent caffeine stimulated bitter-GRNs. In contrast, in GA cockroaches, D-glucose also
stimulated bitter-GRNs and suppressed the responses of sugar-GRNs. Thus, D-glucose is processed as both a
phagostimulant and deterrent in GA cockroaches, and this newly acquired peripheral taste sensitivity
underlies glucose aversion in multiple GA populations. The rapid emergence of this highly adaptive behavior
underscores the plasticity of the sensory system to adapt to rapid environmental change.

ensory systems guide the assessment of

food, habitat, and potential mates, and prominently govern intra- and interspecific inter-

actions. Although great progress has been made

in our understanding of chemosensory processing, especially in insects (1, 2), how chemosensory

Fig. 1. Sensitivities of the GRNs of wild-type (WT) and GA (T164-BC)

cockroaches to various tastants. (A) Side view of the right paraglossa of a
WT male cockroach (left, maxillary and labial palps were removed), and a taste
sensillum used in electrophysiological recordings (right). (B) GRN responses,
showing sample recordings (top) of the same sensillum stimulated sequentially
with fructose, caffeine, and glucose (top); impulse sorting (middle); and hier-

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systems change in response to rapidly changing

environments remains largely unknown. Crossspecies divergence has been well investigated,
particularly in olfactory processes (24). However, identifying the chemosensory mechanisms
that underlie adaptive intraspecific polymorphisms
has been challenging. Among the most important
such polymorphisms are sensory adaptations that
confer behavioral resistance to insecticides (5).
The German cockroach, Blattella germanica,
offers a tractable system to explore mechanisms
of sensory adaptation. Since the mid-1980s, control of this pest has increasingly shifted to baits
that combine an insecticide with various phagostimulants, typically D-glucose (glucose henceforth) and D-fructose (fructose) (6). Within just
several years, cockroach populations evolved a
Department of Entomology and W.M. Keck Center for Behavioral Biology, North Carolina State University, Campus Box
7613, Raleigh, NC 276957613, USA.
*Corresponding author. E-mail: coby@ncsu.edu

archical cluster analysis (bottom). The time bar under each recording indicates
200 ms. (C) Responsiveness of GRNs of WT and GA cockroaches (20 sensilla
each) to 10 tastants. Feeding responses are from fig. S3. Fructose elicited
impulses in GRN1, and caffeine elicited impulses in GRN2 in both strains.
Glucose and related compounds stimulated GRN1 in WT cockroaches and both
GRN1 and GRN2 in GA cockroaches.

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new behavioral traitglucose aversion. Glucoseaverse (GA) cockroaches avoid eating glucosecontaining baits (movies S1 to 4 and fig. S1),
resulting in failure of otherwise highly effective
baits (7). The GA trait is heritable (7, 8), and the
aversive response is robustly evoked by glucose
alone (7, 9). Although growth and reproduction
are slower in GA than in wild-type cockroaches
(10), GA cockroaches outcompete wild-type cockroaches under the strong selection pressure of
glucose-containing baits (7, 11).
We hypothesized that the GA trait could be
encoded by changes in glucose detection. Tastant
detection in insects occurs in peripheral gustatory receptor neurons (GRNs), which are housed
within hairlike sensilla on the mouthparts (12, 13).
The GRNs have modal taste specificity, so in
Drosophila, for example, four GRNs encode four
taste classes: sugar-, bitter-, water- and salt-sensitive
GRNs (13, 14). Each GRN expresses multiple

gustatory receptors (GRs) that recognize tastants

and transduce information about their quality and
strength into neuronal impulses that can be distinguished by their amplitude and duration (15, 16).
As in other animals, tastants that activate sugarGRNs elicit appetitive behavior (13, 17) and
tastants that activate bitter-GRNs drive aversive
behavior (13, 18).
The organization and functions of GRNs in
the German cockroach are poorly understood (19).
We concentrated on glucose-sensitive sensilla on
the paraglossae (Fig. 1A) because the paraglossae
alone can drive glucose acceptance in wild-type
cockroaches and its rejection in GA cockroaches
(9). Analysis of impulse waveforms [Fig. 1B; also
see (20)] and cross-adaptation experiments (fig.
S2) in wild-type cockroaches demonstrated that
glucose-sensitive sensilla contain four distinct GRNs.
Fructose and glucose selectively stimulated GRN1,
whereas caffeine selectively stimulated GRN2.

Fig. 2. Behavioral and electrophysiological responses to six tastants.

(A) Chemical structures of tastants. (B) Dose-feeding responses in WT
(blue) and GA (T164-BC, red) cockroaches motivated to accept phagostimulants but not water (Hungry), or to take both phagostimulants and
water (Hungry and thirsty). Feeding response is the proportion of cockwww.sciencemag.org

GRN3 and GRN4 responded to both sugars and

caffeine. Using a panel of tastants (Fig. 1C and
fig. S3), we established that all tastants that stimulated feeding in wild-type cockroaches also stimulated GRN1 but not GRN2, and all deterrents
stimulated GRN2 but not GRN1. The results indicate that the appetitive and aversive inputs in
wild-type cockroaches segregate by the organization of GRN1 (sugar-GRN) and GRN2 (bitterGRN) at the peripheral sensory level, as in other
insect species (12, 13, 19).
The sugar- and bitter-GRN sensitivities of GA
cockroaches (strain T164-BC) were considerably
different from those of wild-type cockroaches.
Glucose stimulated four rather than only three
types of GRNs (Fig. 1B and fig. S2), corresponding to the sugar-GRN, bitter-GRN, GRN3, and
GRN4 of wild-type cockroaches. Electrophysiological recordings from GA cockroaches with
10 tastants further demonstrated that the bitter-GRN

roaches ingesting the test solution, and the legends indicate sample size.
GA cockroaches rejected glucose and related compounds. (C) The sugarand bitter-GRNs of WT and GA cockroaches respond differentially to six
tastants (mean T SEM). Number of tested sensilla is in parentheses. (*P <
0.05, Students t test).

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Fig. 3. Glucose aversion is elicited by stimulation of bitter-GRNs and

inhibition of sugar-GRNs. (A) Cockroaches were tested with fructose alone
(Fru), fructose mixed with 30 or 300 mmol liter1 glucose (F30G and F300G),
and fructose mixed with 1 or 10 mmol liter1 caffeine (F1C and F10C). Numbers of tested WT and GA (T164-BC) cockroaches are in the legends (in
parentheses). The response to fructose alone is also in Fig. 2B. (B) Sensitivity
of sugar-GRN (top, blue) and bitter-GRN (bottom, red) to fructose alone and to
responded to glucose and all the tastants that elicited aversive behavior (Fig. 1C and fig. S3). We
therefore suggest that glucose and related compounds drive the aversive response in GA cockroaches by stimulating the bitter-GRN, the same
GRN that is stimulated by caffeine in both cockroach strains (Fig. 1C). By contrast, GRN3 and
GRN4 responded without any apparent discrimination among stimuli (Fig. 1C, fig. S4A, and
table S1), suggesting that they do not contribute to the differential discrimination of appetitive and aversive tastants by the two cockroach
strains.
We compared the sensitivities of the sugarand bitter-GRNs in the wild-type and GA strains
with dose-behavioral response studies with six
tastants (Fig. 2A). The two cockroach strains showed
similar behavioral and GRN responses to fructose and caffeine (Fig. 2, B and C), suggesting
that wild-type and GA cockroaches have fundamentally similar gustatory neural networks for appetitive and aversive behaviors. However, glucose
and two related compounds stimulated the bitterGRN in GA cockroaches (Fig. 2, B and C), and
3-O-methyl-D-glucose, which was aversive to both
strains, elicited significantly higher bitter-GRN
responses in GA than in wild-type cockroaches.

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binary mixtures (means T SEM). S, 0.25 mmol liter1 NaCl (control electrolyte);
2F and 4F, 2 and 4 mmol liter1 fructose; 8G and 32G, 8 and 32 mmol liter1
glucose; 0.04C and 0.16C, 0.04 and 0.16 mmol liter1 caffeine. Number of
tested sensilla is in parentheses. The GRN responses to fructose alone were
compared to the responses to binary mixtures (analysis of variance, Dunnetts
test, *P < 0.05). Glucose and caffeine attenuate the feeding response to
fructose in GA cockroaches and depress the sugar-GRN responses.

The results suggest that in wild-type cockroaches,

glucose and related compounds are discriminated
structurally by narrowly tuned receptors on sugarGRNs, eliciting appetitive behavior. In GA cockroaches, by contrast, the expression of a broadly
tuned receptor or multiple narrowly tuned receptors may contribute to the broad acceptance of
glucose and related compounds by bitter-GRNs,
driving aversive behavior.
Sugar-GRNs in GA cockroaches also exhibited a significantly lower response to glucose than
in wild-type cockroaches (Fig. 2C). We tested
whether the sugar-GRNs of GA cockroaches are
less sensitive to glucose, or if their responses are
depressed by the activities of adjacent GRNs.
Complementary behavioral assays and electrophysiological recordings with mixtures of phagostimulants and deterrents revealed that in GA
cockroaches, both glucose and caffeine attenuated the appetitive response to fructose (Fig. 3A
and table S2) and significantly depressed the sugarGRN responses relative to fructose alone (Fig. 3B).
By contrast, in wild-type cockroaches, combining
glucose with fructose increased both the appetitive
response and the electrophysiological responses of
sugar-GRNs compared to fructose alone (Fig. 3B).
These results demonstrate that GA cockroaches

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detect glucose as a genuine deterrent, which

also suppresses sugar-GRN responses, as alkaloids and glucosides do in other insect species
(2123).
How prevalent is this mechanism in glucoseaverse field populations? We screened the feeding
responses of 19 field-collected populations and
found seven populations with GA cockroaches
(Fig. 4A). Two of these strains were used in behavioral and GRN dose-response studies. Although
both were less GA than the lab-selected strains
(Fig. 4B and table S2), in both strains glucose
stimulated the bitter-GRN (Fig. 4C) and depressed
the sugar-GRN (table S1). In four GA strains, the
behavioral feeding responses negatively correlated with bitter-GRN responses (Fig. 4D and table
S3). The wild-type and field-collected strains did
not differ in GRN sensitivities for both fructose
and caffeine (fig. S5 and table S1), confirming
that a similar mechanism gave rise to glucose aversion in multiple cockroach populations.
Most natural genetic polymorphisms in taste
receptors modify behavioral responses over a finite range, from exquisite sensitivity to complete
insensitivity to a particular tastant [e.g., (24)]. In
bait-selected cockroach populations, however, the
modal specificity of glucose has been dramatically

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Fig. 4. Glucose stimulates bitter-GRNs in field-collected cockroaches.

(A) Behavioral assays showing 7 of 19 field-collected populations with some
GA cockroaches. (B) Dose-feeding responses to glucose in four GA strains, with
the number of tested cockroaches in parentheses. T164-BC response to
glucose is also shown in Fig. 2B, and the median effective concentration (EC50)

transformed from sweet and highly phagostimulatory to bitter and highly deterrent. Generally,
bitter-GRNs of insects coexpress a large number
of GRs (18, 25) and are therefore broadly tuned
to respond to various deterrents (18, 21, 22). The
coexpression patterns of GRs ultimately account
for the unique sensitivity of bitter-GRNs and their
capacity to selectively respond to specific deterrents (18). Our electrophysiological studies with
GA cockroaches suggest two major hypotheses:
One or more mutations have either (i) modified
the structure of GRs on the bitter-GRN to accept
glucose and/or (ii) caused the misexpression of
native glucose GRs on the bitter-GRN. The structureactivity studies tentatively support the former hypothesis that the glucose-sensitive GRs on bitter-GRNs
are differently tuned from the native glucose GRs
on sugar-GRNs, because wild-type and GA
cockroaches responded differentlyboth behaviorally and with GRN responsesto changes in
the chemical structures of glucose and related
compounds.
Our results show that by recruiting glucose
and related sugars as bitter-GRN ligands, a gainof-function adaptation has emerged, expressing
glucose-aversion as a novel behavior that offers
protection against toxic baits. The change in valence of glucose, without compromising the exquisite sensitivity of the gustatory system to glucose,
highlights the specificity of this adaptive change.
Moreover, the aversion to glucose is further amplified by a preexisting inhibition of sugar-GRN
responses by deterrents. Glucose aversion is a

for each strain is in table S2. (C) Dose-GRN responses to glucose in WT and
four GA strains (mean impulse frequency T SEM, with number of tested sensilla
in parentheses). (D) Feeding responses [from (B)] and GRN2 responses [from
(C)] at similar glucose concentrations are negatively correlated (r, Pearson's
correlation coefficient, P < 0.001, table S3).

clear example of a chemosensory gain-of-function

adaptation that confers behavioral resistance to
anthropogenic pressures, protecting the German
cockroach from insecticides.
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Acknowledgments: We thank R. Santangelo, V. Knowlton,
A. Ernst, J. Mahaffey, R. Anholt, N. Bao, D. Bieman, and
D. Mukha for equipment and cockroach collections and
T. Tanimura, F. Marion-Poll, A. A. Dahanukar, and three
anonymous reviewers for constructive comments. This
research was supported in part by NSF (IOS-1052238) and
U.S. Department of Housing and Urban Development
(NCHHU0001-11) awards to C.S. and by the Blanton
J. Whitmire Endowment at North Carolina State University.
Data associated with this manuscript have been archived in
DRYAD Digital Repository (http://datadryad.org/).

Supplementary Materials
www.sciencemag.org/cgi/content/full/340/6135/972/DC1
Materials and Methods
Figs. S1 to S5
Tables S1 to S3
References (2628)
Movies S1 to S4
7 January 2013; accepted 26 March 2013
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